Chromatography- Separation of mixtures CHEM 212. What is solvent extraction and what is it commonly used for?
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1 Chromatography- Separation of mixtures CHEM 212 What is solvent extraction and what is it commonly used for? How does solvent extraction work? Write the partitioning coefficient for the following reaction: Sphase1 ó Sphase2 Write the equation for calculating the fraction remaining in phase 1 after n extractions. What is the distribution coefficient? How is it different from K? Which solvent is preferred by the charged and uncharged species? Charged Uncharged Write the equation for D in terms of Ka for an acid and base solute Base Acid Write the equation for D for metal extraction using a chelator in terms of equilibrium constants.
2 Describe in general terms what chromatography does and how it works. Define the following terms in words and with an equation and/or units, if applicable. Term Symbol Equation Description Retention time tr Adjusted retention time Volume flow rate t r Linear flow rate Relative retention Partition coefficient retention factor α K k Peak width FWHM W½ Resolution Rs Define the retention factor in words. Relate the relative retention time to the partition coefficient Relate relative retention with the partition coefficients
3 Describe the relationship between tr, tm, and tr with respect to the interaction of the mobile phase, stationary phase, and solute. Label tr, tm, and tr in the diagram below. Describe the basis of separation for each of the types of chromatography described below. Absorption Chromatography Partitioning Chromatography Ion Chromatography Size Exclusion Chromatography Affinity Chromatography
4 Chromatography- Separation of mixtures 1. Sample dissolved in mobile phase 2. Mobile phase passes over stationary phase 3. Analyte molecules partition between mobile and stationary phase 4. Different solutes will elute at different times Separation of mixtures Resolution of separations State how resolution is calculated? What is considered quantitative separation? Better separations require either: 1. increase in rate of separations 2. decrease in rate of band spreading Define diffusion in words. Why does it happen? State the equation for the diffusion coefficient What is the relationship between bandwidth and retention time?
5 What does a theoretical plate represent? How is plate height calculated? State the equation and define each variable State the equation for calculating the number of plates using full width half max (w1/2) What is the relationship between bandwidth and plate height? What factors affect resolution and how are they each related to resolution? Comparing chromatographic techniques TLC GC HPLC Stationary phase Silica gel on glass plate Capillary with liquid or solid phase Silica beads- often derivitized with C- 18, etc Mobile phase solvent Gas, usually He Organic or polar solvent or mixture of solvents Column length ~10cm m ~5-25 cm Pressure ambient low Up to 1,- 000 bar Temperature ambient variable Variable
6 High Performance Liquid Chromatography (HPLC) Draw an HPLC instrument diagram: HPLC pressure- up to 1,000 bar (~ bar normal) Column length ~10-25 cm with ~5um C- 18 derivativized silica particles Solvent: Isocratic elution Gradient elution Detectors: UV- Vis ELSD Mass Spectrometry
7 Draw a GC instrument diagram: Gas Chromatography (GC) State the function of each of the components of a GC: Carrier gas Sample injection Headspace injection- Liquid injection- Injector chamber- Column Detector options
8 Creative Project lite- be prepared to present your application and technique in class. 1. Find an application of GC- MS or HPLC you are interested in. Describe the problem and how mass spectrometry is used to address it. Learn about the instrument used in your application and diagram it below. Be sure to include information about the instrument parameters. Be prepared to diagram the instrument and describe each component.
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